慢病毒介导的靶向沉默信息调节因子1（SIRT1）的RNA干扰对肝癌细胞生长及凋亡的影响  

Effect of Lentivirus-Mediated RNA Interference Targeting SIRT1 on Cell Growth and Apoptosis in Hepatocelluar Carcinoma

作　　者：任吉华[1] 宋春丽[1] 陶颖[1] 陈娟[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学重点实验室,重庆市400016

出　　处：《医学分子生物学杂志》2012年第3期178-182,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金（N0.81171628）

摘　　要：目的探讨慢病毒介导的靶向SIRTlshRNA对肝癌细胞生长和凋亡的影响。方法Western印迹分析SIRT1在多个肝癌细胞系中的表达；通过慢病毒介导的shRNA干扰技术靶向沉默SIRT1的表达，并通过Western印迹验证SIRTl基因的沉默效果。台盼蓝排斥实验分析SIRT1基因沉默对肝癌细胞生长的影响；流式细胞术和Western印迹检测PARP蛋白的剪切物观察细胞凋亡状态。结果SIRT1在多个肝癌细胞系中表达水平明显上调；慢病毒介导的shRNA能显著抑制细胞中SIRT1的表达。流式细胞术及Western印迹结果均显示SIRT1表达沉默显著诱导了肝癌细胞的凋亡。结论慢病毒介导的靶向SIRT1shRNA显著地抑制SIRT1的表达；SIRT1基因沉默抑制肝癌细胞生长并促进了细胞凋亡。Objective To investigate the effect of SIRT1 downregulation on cell proliferation and apoptosis. Methods The expression of SIRT1 in the HCC cell lines was analyzed by western blot. SIRT1 expression was downregulated by lentivirus-mediated shRNA targeting SIRT1. SIRT1 protein level was measured by western blot. The effect of SIRT1 gene silencing on HCC cell prolifer- ation was measured by trypan blue exclusion assay. The apoptosis was observed by flow cytometry and western analysis of PARP cleavage. Results SIRT1 was overexpressed in the HCC cell lines examined. Knockdown of SIRT1 inhibited the proliferation of HCC cells and induced the apoptosis, as determined by flow cytometry and western blot results. Conclusion SIRT1 is overexpressed in the HCC cell lines. Knockdown of the overexpressed SIRT1 can inhibit cell proliferation and induce apoptosis of the HCC cells.

关 键 词：原发性肝癌 SIRT1 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]